College of Animal Science and Technology, Heilongjiang Bayi Agricultural University, Daqing 163319, China;Laboratory Animal Center of the Academy of Military Medical Sciences, Beijing 100071 在期刊界中查找 在百度中查找 在本站中查找
Laboratory Animal Center of the Academy of Military Medical Sciences, Beijing 100071;College of Animal Science and Veterinary Medicine, Shenyang Agricultural University, Shenyang 110161 在期刊界中查找 在百度中查找 在本站中查找
Laboratory Animal Center of the Academy of Military Medical Sciences, Beijing 100071;College of Animal Science and Veterinary Medicine, Jilin University, Changchun 130062 在期刊界中查找 在百度中查找 在本站中查找
Objective To construct recombinant eukaryotic expression vectors of Beagles ERβ1293, and identify the expression and localization of ERβ1293 in the human embryonic renal HEK293T cells by Western blotting and indirect immunofluorecent assay.Methods The ERβ1293 gene was amplified by PCR using pEGFP-N1-ERβ1293 as a template. The recombinant eukaryotic expression vector of pcDNA3.1-Myc-ERβ1293 was constructed, and transfected it into HEK293T cells. The expression of pcDNA3.1-Myc-ERβ1293 was detected by Western blot, and its localization was detected by indirect immunofluorecence assay. Results The expression of pcDNA3.1-Myc-ERβ1293 was constructed, and successfully identified in the HEK293T cells using Western blotting, and detected only in the cytoplasm using IF by laser scanning confocal microscopy. Conclusions In the previous experiment, the coding sequence of Beagles ERβ1593 splice variants has been obtained, which is lack of exon 4, and its ligand binding ability is reduced or disappeared, so that the intracellular localization of ERβ1293 codeing protein is changed.